水通道蛋白AQP2在原发性肝细胞肝癌多柔比星耐药机制中的作用研究

摘要

目的 探索原发性肝细胞肝癌(HCC)多柔比星(doxorubicin,DOX)耐药的机制,为进一步提高HCC患者介入治疗的疗效提供前期探索.方法 应用Western blotting方法检测DOX处理后的上皮间质化转变(EMT)的相关蛋白E-Cadherin和Vimentin表达情况,使用流式细胞仪观察细胞干性标记CD133的变化情况,使用基因芯片技术观察DOX处理后的细胞机制变化.结果 DOX能诱导HCC细胞发生上皮间质化转变,并且能提升HCC细胞CD133的比例(3.5%vs 35.5%,P<0.05);基因芯片提示HCC细胞内水通道蛋白AQP2基因显著上调;敲降AQP2基因后,能显著减弱DOX诱导HCC细胞产生EMT的效果.结论 水通道蛋白AQP2在肝细胞肝癌多柔比星耐药机制中具有重要作用,降低AQP2的表达可以减弱多柔比星诱导肝癌细胞上皮间质化转变的效果.%Objective To investigate the mechanisms of doxorubicin resistance in hepatocellular carcinoma (HCC), and to provide preliminary exploration on improving the effect of transhepatic arterial chemotherapy and embolization (TACE) in HCC patients.Methods The expression of epithelial-mesenchymal transition (EMT) related proteins, E-Cadherin and Vimentin were detected by Western blotting. Flow cytometry was used to evalu-ate the level of CD133. Furthermore, the precise mechanisms under doxorubicin treatment was analyzed by mi-croarray.Results Doxorubicin could significantly induce EMT in HCC cells. Besides, the expression of CD133 was also increased (3.5%vs 35.5%,P<0.05). The microarray data suggested that Aquaporins 2 (AQP2) increased most dramatically among AQPs under doxorubicin treatment. In addition, knockdown of AQP2 significantly at-tenuated the induction of EMT.Conclusion AQP2 plays an important role in doxorubicin resistance in HCC. Knockdown of AQP2 may reduce the doxorubicin-induced EMT.